This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project is designed to investigate the diversification pattern of the self-incompatibility locus (S-locus) in the poppy genus, apaver. Self-incompatibility (SI) is a genetic system that allows plants to avoid self-fertilization. Self-recognition loci, such as the self-incompatibility loci in plants are stunningly diverse, with both large numbers of functionally different alleles, and deep divergence among alleles as a result of negative frequency dependent selection. Our previous work has used degenerate PCR to discover new S-alleles. However, this approach has been inefficient and has likely introduced bias in the alleles that we have discovered to date due to low sequence conservation among different alleles in the S-locus. A more efficient, less biased approach is required for an accurate picture of molecular population genetic diversity at this locus. We propose to test whether next-generation sequencing (NGS) technology can be employed for improved discovery of new S-locus alleles.